Molecular Plant Breeding 2024, Vol.15, No.1, 15-26 http://genbreedpublisher.com/index.php/mpb 21 4.2 Examples and advantages of genome editing breeding Genome editing is another application area with tremendous potential in Breeding 4.0. One of the landmark papers in genome editing technology was published in 2014 in the journal “Human Molecular Genetics”. This research described the CRISPR-Cas9 system composed of the nonspecific Cas9 nuclease and a set of programmable specific sequence CRISPR RNA (crRNA), which guides Cas9 to cleave DNA, thereby generating double-strand breaks at the target site. Subsequent cellular DNA repair processes lead to the desired insertions, deletions, or replacements at the target site (Zhang et al., 2014). Through CRISPR-Cas9 technology, researchers can precisely modify specific gene sequences in the genome, achieving accurate control over traits. The breakthrough in this technology has revolutionized the field of genome editing, providing new tools and methods for Breeding 4.0. By utilizing gene editing technologies like CRISPR-Cas9, breeders can precisely modify specific gene sequences in the crop genome, achieving accurate control and improvement of traits. Gene editing can be employed to enhance disease resistance, improve quality characteristics, and increase yield in crops. For instance, Veillet et al. (2019) successfully achieved efficient editing of target bases by targeting the acetyl lactate synthase (ALS) genes of tomatoes and potatoes, by using the CRISPR/Cas9 gene editing tool, creating herbicide-resistant plants. And 12.9% of tomatoes and 10% of potatoes in the first generation of plants do not contain genetically modified ingredients. The study by Yasumoto et al. (2020) demonstrated the application of gene editing in potato breeding by using TALENs and CRISPR-Cas9 technologies. Precise editing of the potato genome was achieved through transient TALEN expression via Agrobacterium infection, holding significant implications for crop breeding. Kawall’s research (2021) discussed methods to accelerate modern plant breeding by using gene editing technologies. By altering specific features of rapeseed through gene editing, such as increasing yield and modifying nutritional components, the study showcased the enormous potential of this technology in plant breeding. Nerkar et al. (2022) emphasized the powerful potential of gene editing technology in precisely modifying specific loci in the crop genome, which is a long-term goal for crop breeders. The study also discussed the regulatory approval status of gene-edited crops and the prospects of developing high-yielding stress-resistant crops through precise gene editing. 4.3 The contribution of Breeding 4.0 to the sustainable development of agriculture The application of Breeding 4.0 has made positive contributions to the sustainable development of agriculture. The use of innovative technologies such as gene editing and transgenic breeding can enhance crop resistance and adaptability, reducing dependence on pesticides and fertilizers, thereby minimizing the negative environmental impact of agriculture. Additionally, Breeding 4.0 can address the growing demand for food by increasing crop yields, improving quality, and enhancing stress tolerance, providing a more sustainable food supply. The innovations and progress in Breeding 4.0 offer new pathways and possibilities for achieving green, efficient, and sustainable agricultural development. Through the development of applications such as transgenic breeding and gene editing, Breeding 4.0 can offer more choices and solutions for agricultural production. These applications demonstrate significant potential in enhancing crop quality, increasing resistance, improving yields, and making important contributions to the sustainable development of agriculture. 5 Challenges and Prospects of Breeding 4.0 5.1 Considerations of ethical, legal, and social issues Breeding 4.0 introduces new genetic information integration and editing technologies, triggering a series of considerations regarding ethical, legal, and social issues. For instance, transgenic breeding has sparked controversies related to food safety, environmental impact, and intellectual property rights. The application of gene editing technology also involves ethical concerns and considerations of societal acceptance. Whelan and Lema (2017) explored the application of gene editing technology in plant breeding, with a particular focus on whether these technologies should be considered a form of conventional breeding or be subject to regulations governing genetically modified crops. The research indicates that policymakers should consider safety
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